1. Field
The present disclosure relates to a polynucleotides used for the detection, analysis, and amplification of target nucleic acids.
2. Description of the Related Art
Samples obtained in the medical field are usually small volume samples and present as mixtures containing various kinds of nucleic acids. As a result, it is generally necessary to amplify target nucleic acids present in a sample to obtain a sufficient amount of the target for accurate analysis.
Current methods of target nucleic acid amplification utilizing primers may undesirably produce non-specific reactants through the formation of a primer dimer during the amplification process. Non-specific reactants are problematic in methods for nucleic acid amplification in that the target nucleic acids are not amplified and/or the sensitivity of the target nucleic acids to the amplification is lowered. In polymerase chain reaction (PCR), since the reaction occurs at high temperatures, the specificity of primers used to amplify target nucleic acids is high. However, when a mixture for PCR is manufactured at room temperature, for example, a temperature in a range of about 20° C. to about 25° C., a primer dimer can be easily formed. In order to inhibit such primer dimer formation, a method of ‘hot start’ nucleic acid amplification has been developed, which impedes the amplification until the temperature goes up. For example, a method of hot start nucleic acid amplification may include an anti-polymerase antibody that non-covalently binds a polymerase to inactivate it at low temperatures. However, since antibodies are used in such a method, the cost is high and there may be problems of mammalian genomic DNA contamination. Additionally, hot start methods present the difficulty of determining whether all the polymerase and anti-polymerase antibody are bound.
Accordingly, there remains a need for the development of a cost effective and improved method of hot start nucleic acid amplification that inhibits the primer dimer formation without an antibody when preparing a PCR mixture at room temperature.